Nitro blue tetrazolium

Nitro blue tetrazolium is a chemical compound composed of

two tetrazole moieties. It is used in immunology for sensitive
detection of alkaline phosphatase (with BCIP). NBT serves as
the oxidant and BCIP is the AP-substrate (and gives also dark
blue dye).
Clinical significance
In immunohistochemistry the alkaline phosphatase is often used
as a marker, conjugated to an antibody. The colored product can
either be of the NBT/BCIP reaction reveals where the antibody is
bound, or can be used inimmunofluorescence.[1]
The NBT/BCIP reaction is also used for
colorimetric/spectrophotometric activity
assays of oxidoreductases. One application is in activity
stains in gel electrophoresis, such as with the
mitochondrial electron transport chaincomplexes.[2]
Nitro blue tetrazolium is used in a diagnostic test,[3] particularly
for chronic granulomatous disease and other diseases of
phagocyte function. When there is an NADPH oxidase defect, the
phagocyte is unable to make reactive oxygen species or radicals
required for bacterial killing. As a result, bacteria may thrive within
the phagocyte. The higher the blue score, the better the cell is at
producing reactive oxygen species.[4]
References[edit]
1.

Jump up^ Trinh le A, McCutchen MD, Bonner-Fraser

M, Fraser SE, Bumm LA, McCauley DW (June 2007).
"Fluorescent in situ hybridization employing the
conventional NBT/BCIP chromogenic
stain". BioTechniques 42 (6): 756
9.doi:10.2144/000112476. PMID 17612300.

2.

Jump up^ Nisimoto Y, Wilson E, Heyl BL, Lambeth JD

(5 January 1986). "NADH dehydrogenase from bovine
1

Nitro blue tetrazolium

neutrophil membranes. Purification and properties". J. Biol.
Chem. 261 (1): 28590. PMID 3941077.
3.

Jump up^ Freeman, R; King B (October

1972). "Technique for the performance of the nitro-blue
tetrazolium (NBT) test". Journal of Clinical
Pathology 25 (10): 912
914. doi:10.1136/jcp.25.10.912. PMC 477548. PMID 411900
8.

4.

Jump up^ Nathan DG, Baehner RL, Weaver DK

(October 1969). "Failure of nitro blue tetrazolium reduction
in the phagocytic vacuoles of leukocytes in chronic
granulomatous disease". J. Clin. Invest. 48 (10): 1895
904. doi:10.1172/JCI106156.PMC 322426. PMID 5387730.

Nitro blue tetrazolium

5-Bromo-4-chloro-3-indolyl phosphate
From Wikipedia, the free encyclopedia
(Redirected from BCIP)

BCIP

Names
IUPAC name
(5-bromo-4-chloro-1H-indol-3-yl) dihydrogen phosphate
Other names
BCIP
Identifiers
CAS Number

38404-93-2

ChemSpider

58873

InChI[show]

Nitro blue tetrazolium

Jmol interactive 3D

Image
Image

PubChem

65409

SMILES[show]
Properties
Chemical formula

C8H6BrClNO4P

Molar mass

326.47 gmol1

Appearance

Colorless

Solubility in water

soluble in water (sodium salt)

Except where otherwise noted, data are given for materials in

their standard state (at 25 C [77 F], 100 kPa).
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Infobox references

5-Bromo-4-chloro-3-indolyl phosphate (BCIP, X-phosphate, XP) is an artificial

chromogenic substrate used for the sensitive colorimetric detection of alkaline phosphatase activity.
It is, for example, used in immunoblotting, in situ hybridization, and immunohistochemistry, often in
combination with nitro blue tetrazolium chloride (NBT).[1][2] 5-bromo-4-chloro-3-indoxyl is oxidized by
atmospheric oxygen to form the blue dye 5,5-dibromo-4,4-dichloro-indigo. It is also oxidized by
nitroblue tetrazolium (NBT), which forms an insoluble dark blue diformazan precipitate after
reduction. Alkaline phosphatase is commonly conjugated to secondary antibodies.

References[edit]
1. Jump up^ http://www.sigmaaldrich.com/catalog/search/ProductDetail/SIGMA/B1026
2. Jump up^ Alkaline phosphatase hydrolyses BCIP to 5-bromo-4-chloro-3-indoxyl<JP Horwitz J. Med.
Chem., 1966, 9 (3), pp 447447 and inorganic phosphate